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The purpose of this research was to investigate the effect of toluidine blue (TB) mediated photodynamic therapy (PDT) on the inhibition of lipopolysaccharide (LPS)-induced inflammation in rat gingival fibroblasts through in vitro experiments. Rat gingival fibroblasts were divided into five groups: (1) control, (2) LPS treatment, (3) laser treatment, (4) TB treatment (1.0 µg/mL), and (5) PDT treatment (TB plus laser irradiation at 320 mW/cm2 for 240 s). After 24 h, cell growth activity was measured using MTT assay. The levels of receptor activator for nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG) in the cell culture supernatant were measured using enzyme-linked immunosorbent assay (ELISA). Nuclear proteins were extracted and the phosphorylation levels of phosphorylated nuclear factor-κB/p65 (p-p65) and phosphorylated inhibitor of nuclear factor-κB (p-IκBα) were determined using Western Blot. MTT results showed no significant difference in cell viability between the groups (P > 0.05). After LPS induction, OPG expression decreased, RANKL expression increased, and the OPG/RANKL ratio decreased, which was different from the control group (P < 0.05). After PDT treatment, OPG expression increased, RANKL expression decreased (P < 0.05), and the OPG/RANKL ratio increased (P < 0.05). Compared to the control group, there was no significant difference in OPG and RANKL expression or the OPG/RANKL ratio (P > 0.05). The activation of NF-κB was closely related to the phosphorylation levels of p-p65 and p-IκBα. LPS significantly up-regulated p-p65 and p-IκBα expression (P < 0.05), while PDT treatment decreased their phosphorylation levels (P < 0.05). TB-PDT treatment can inhibit NF-κB signaling pathway activation, decrease RANKL and OPG expression, and reduce the OPG/RANKL ratio, thereby reducing inflammation and playing a role in periodontitis treatment. [ABSTRACT FROM AUTHOR]

In this study, dead Bacillus niacini microorganisms were used to support the immobilization of magnetic iron nanoparticles, creating a magnetic nano-biosorbent for wastewater treatment through magnetic separation. Magnetic nano-biosorbent was characterized via scanning electron microscopy, transmission electron microscopy, energy-dispersive X-ray spectroscopy, Fourier transform infrared spectrophotometry, Brunauer–Emmett–Teller analysis, and vibration sample magnetometry techniques. The laser particle sizer confirmed a uniform distribution in the particle agglomerate sizes of magnetic iron nanoparticles and magnetic nano-biosorbent, affirming successful composite formation. Energy-dispersive X-ray spectroscopy confirmed Bacillus niacini specific elements, and Fourier transform infrared spectrophotometry indicated effective Bacillus niacini coating onto magnetic iron nanoparticles. Magnetic nano-biosorbent's efficacy for toluidine blue and lead (II) removal, considering pH, contact time, magnetic nano-biosorbent dosage, and initial pollutant concentrations, was assessed. Langmuir isotherms described toluidine blue and lead (II) biosorption optimally. Kinetic data matched the pseudo-first-order and pseudo-second-order models, implying multiple biosorption mechanisms. Magnetic nano-biosorbent displayed a biosorption capacity of 66.52 ± 0.68 mg/g for lead (II) and 82.88 ± 0.79 mg/g for toluidine blue. Reusability tests showed effective reuse for up to five cycles. The magnetic nano-biosorbent presents significant potential for wastewater treatment due to its high biosorption capacity, efficient removal, and cost-effective synthesis. [ABSTRACT FROM AUTHOR]

This study investigates the synthesis, characterization, and application of ZnO/SnO2 nanocomposite (ZnO/SnO2 NC), focusing on their photocatalytic and antioxidant properties. The sol-gel technique was utilized for the synthesis of the ZnO/SnO2 NC. Subsequent characterization was performed using X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, and UV-visible (UV-Vis) spectrophotometry. The XRD analysis indicated crystallite dimensions of 15 nm for ZnO/SnO2 and 21 nm for pure ZnO NPs. The photocatalytic efficiency was assessed by the degradation of Toluidine Blue (TB) and m-Toluidine Blue (m-TB) dyes. The ZnO/SnO2 NC demonstrated degradation rates of 99% for both TB and m-TB within 120 and 140 min of sunlight exposure, respectively. Furthermore, the scavenging activity of superoxide radicals was evaluated, with the ZnO/SnO2 NC exhibiting an Half-maximal inhibitory concentration (IC50) value of 48.12 µg∙mL− 1, compared to 60.03 µg∙mL− 1 for pure ZnO. This research highlights the potential of ZnO/SnO2 NC in environmental remediation and biomedical applications. [ABSTRACT FROM AUTHOR]

The development of nanomaterials has been speedily established in recent years, yet nanoparticles synthesized by traditional methods suffer unacceptable toxicity and the sustainability of the procedure for synthesizing such nanoparticles is inadequate. Consequently, green biosynthesis, which employs biopolymers, is gaining attraction as an environmentally sound alternative to less sustainable approaches. Chitosan-encapsulated nanoparticles exhibit exceptional antibacterial properties, offering a wide range of uses. Chitosan, obtained from shrimp shells, aided in the environmentally friendly synthesis of high-purity zinc oxide nanoparticles (ZnO NPs) with desirable features such as the extraction yield (41%), the deacetylation (88%), and the crystallinity index (74.54%). The particle size of ZnO NPs was 12 nm, while that of chitosan–ZnO NPs was 21 nm, and the bandgap energies of these nanomaterials were 3.98 and 3.48, respectively. The strong antibacterial action was demonstrated by ZnO NPs, chitosan–ZnO NPs, and chitosan–ZnO/PVP, particularly against Gram-positive bacteria, making them appropriate for therapeutic use. The photocatalytic degradation abilities were also assessed for all nanoparticles. At a concentration of 6 × 10–5 M, chitosan removed 90.5% of the methylene blue (MB) dye, ZnO NPs removed 97.4%, chitosan-coated ZnO NPs removed 99.6%, while chitosan–ZnO/PVP removed 100%. In the case of toluidine blue (TB), at a concentration of 4 × 10–3 M, the respective efficiencies were 96.8%, 96.8%, 99.5%, and 100%, respectively. Evaluation of radical scavenger activity revealed increased scavenging of ABTS and DPPH radicals by chitosan–ZnO/PVP compared to individual zinc oxide or chitosan–ZnO, where the IC50 results were 0.059, 0.092, 0.079 mg/mL, respectively, in the ABTS test, and 0.095, 0.083, 0.061, and 0.064 mg/mL in the DPPH test, respectively. Moreover, in silico toxicity studies were conducted to predict the organ-specific toxicity through ProTox II software. The obtained results suggest the probable safety and the absence of organ-specific toxicity with all the tested samples. [ABSTRACT FROM AUTHOR]

Objective and design: Mast cells (MCs), as the fastest immune responders, play a critical role in the progression of neuroinflammation-related diseases, especially in depression. Quercetin (Que) and kaempferol (Kae), as two major diet-derived flavonoids, inhibit MC activation and exhibit significant antidepressant effect due to their anti-inflammatory capacity. The study aimed to explore the mechanisms of inhibitory effect of Que and Kae on MC activation, and whether Que and Kae suppress hippocampal mast cell activation in LPS-induced depressive mice. Subjects and treatment: In vitro assays, human mast cells (HMC-1) were pretreated with Que or Kae for 1 h, then stimulated by phorbol 12-myristate 13-acetate (PMA) and 2,5-di-t-butyl-1,4-benzohydroquinone (tBHQ) for 3 h or 12 h. In vivo assays, Que or Kae was administered by oral gavage once daily for 14 days and then lipopolysaccharide (LPS) intraperitoneally injection to induce depressive behaviors. Methods: The secretion and expression of TNF-α were determined by ELISA and Western blotting. The nuclear factor of activated T cells (NFAT) transcriptional activity was measured in HMC-1 stably expressing NFAT luciferase reporter gene. Nuclear translocation of NFATc2 was detected by nuclear protein extraction and also was fluorescently detected in HMC-1 stably expressing eGFP-NFATc2. We used Ca2+ imaging to evaluate changes of store-operated calcium entry (SOCE) in HMC-1 stably expressing fluorescent Ca2+ indicator jGCamP7s. Molecular docking was used to assess interaction between the Que or Kae and calcium release-activated calcium modulator (ORAI). The hippocampal mast cell accumulation and activation were detected by toluidine blue staining and immunohistochemistry with β-tryptase. Results: In vitro assays of HMC-1 activated by PtBHQ (PMA and tBHQ), Que and Kae significantly decreased expression and secretion of TNF-α. Moreover, NFAT transcriptional activity and nuclear translocation of NFATc2 were remarkably inhibited by Que and Kae. In addition, the Ca2+ influx mediated by SOCE was suppressed by Que, Kae and the YM58483 (ORAI inhibitor), respectively. Importantly, the combination of YM58483 with Que or Kae had no additive effect on the inhibition of SOCE. The molecular docking also showed that Que and Kae both exhibit high binding affinities with ORAI at the same binding site as YM58483. In vivo assays, Que and Kae significantly reversed LPS-induced depression-like behaviors in mice, and inhibited hippocampal mast cell activation in LPS-induced depressive mice. Conclusions: Our results indicated that suppression of SOCE/NFATc2 pathway-mediated by ORAI channels may be the mechanism of inhibitory effect of Que and Kae on MC activation, and also suggested Que and Kae may exert the antidepressant effect through suppressing hippocampal mast cell activation. [ABSTRACT FROM AUTHOR]

Dyes as one of the majority critical environmental pollutants from industrial wastewaters, have caused many concerns for researchers. In this research an effective photocatalyst based on La2O3-SiO2 nanocomposite was prepared and evaluated for degradation of toluidine blue (TB) dye. The prepared photocatalyst was illustrated impressive performance in degradation of TB equal to 97% by visible light irritation. Accuracy of synthesized nanocomposite was studied by some technics including FE-SEM, FT-IR, EDX and mapping analysis. Also the effective parameters in degradation percent of TB such as pH, temperature, time, concentration and the amount of nanocomposite were investigated and optimized equal to amounts of 8.0, 25℃, 80 min, 3.0 µg/mL and 0.04 mg respectively. [ABSTRACT FROM AUTHOR]

The development of effective solid adsorbents is necessary to remove organic pollutants from wastewater in response to the global water crisis. The present study investigates the adsorption behavior of dye pollutants on a 1, 3, 6, 8-tetrakis (4formylphenyl) pyrene (TFPPy) for the self-assembly of an innovative three-dimensional (3D) covalent organic frameworks (COFs). Molecular dynamics (MD) and the well-tempered metadynamics simulation were used to evaluate the ability of COFs to adsorb seven types of dye pollutants (DPs). These include methyl orange (MO), gentian violet (GV), brilliant green (BG), tolonium chloride (TC), methylene blue (MB), anionic congo red (CR), and rhodamine B (Rh B). To examine this process, several descriptors are calculated from the simulation trajectories, including interaction energies, root-mean-square deviation, radial distribution function, solvent-accessible surface area, mean square displacement, and the number of hydrogen bonds to figure out the process. After analyzing the data, it was found that the MO/COFs system displayed exceptional stability. It was confirmed by a significant value of −448.54 KJmol−1 for total interaction energy. The density profile of dye molecules in the MO and Rh B-COFs systems exhibits the highest peaks at approximately 325.79, and 26.75 kg/m³, respectively. The free energy values for the DPs/COFs complexes at their global minima reached about, −512.127, −435.603, −381.244, −300.313, and −283.230 kJmol-1 for MO/GV/BG/TC/ and Rh B with the COFs, respectively. The nature of the design of TFFPy-COF's unique, ultrafast kinetics, high adsorption capacity, and good reusability endow them with great potential for removing dyes in an aqueous environment. [ABSTRACT FROM AUTHOR]

Cancer antigen 125 (CA125) is the most common biomarker used to diagnose and monitor ovarian cancer progression for the last four decades, and precise detection of its levels in blood serum is crucial. In this work, label-free impedimetric CA125 immunosensors were fabricated by using screen-printed carbon electrodes modified with poly toluidine blue (PTB) (in deep eutectic solvent)/gold nanoparticles (AuNP) for the sensitive, environmentally friendly, economical, and practical analysis of CA125. The materials of PTBDES and AuNP were characterized by Fourier Transform Infrared (FT-IR), Scanning Electron Microscope (FE-SEM), and X-ray Diffraction (XRD). The analysis of the CA125 was performed by electrochemical impedance spectroscopy and the developed immunosensor. The immunosensor's repeatability, reproducibility, reusability, selectivity, and storage stability were examined. The developed label-free immunosensor allowed the determination of CA125 in fast, good repeatability and a low limit of detection (1.20 pg mL−1) in the linear range of 5–100 pg mL−1. The stable surface of the fabricated immunosensor was successfully regenerated ten times. The application of immunosensors in commercial human blood serum was performed, and good recoveries were achieved. The disposable label-free impedimetric CA125 immunosensor developed for the rapid and practical detection of CA125 is a candidate for use in point-of-care tests in clinical applications of ovarian cancer. [ABSTRACT FROM AUTHOR]

Aims: To evaluate the effectiveness of toluidine blue for obtaining safe margins in oral squamous cell carcinoma. Materials and methods: Intra-operatively irrigation of the lesion and its surrounding areas was done with toluidine blue solution for 20 s. Followed by irrigation with 1% acetic acid to remove all the mechanically retained stain. The unstained margins were demarcated using incision placed by no 15 BP blade. The lesion was resected with a safe margin of 1 cm away from the stained tissue. Neck dissection was done according to the nodal status. The tumor along with the resected margins was sent for histopathological examination. Statistical analysis was performed to calculate specificity of the vital stain. The cross tabulation between epithelium of the stained and unstained margins was done and subjected to Chi-square test to calculate the significance. Results: The toluidine blue vital stain has a sensitivity of 93.33%. Out of 15 cases, 2 patients recorded positive excision margins leading to recurrence at primary site; 1 patients recorded positive excision margins leading to recurrence at secondary site; 1 patient recorded free excision margins but had recurrence at secondary site; remaining 11 patients recorded free excision margins and did not have recurrence. Conclusion: Vital staining with toluidine blue is concluded to be specific in demarcating the dysplastic tissue adjacent to the carcinomatous lesion, which when excised along with the adjacent dysplastic tissue leads to a decrease in the recurrence in oral squamous cell carcinoma cases. Furthermore, it is inexpensive, easily available and does not add significantly to the operating time. Moreover, it provides a gross visualization of dysplasia surrounding the lesion especially in cases where in the margins are not well defined. Hence, toluidine blue can be a useful and inexpensive adjunct to identify margins intra-operatively in the current scenario where intra-operative frozen sections are not available. [ABSTRACT FROM AUTHOR]

Plant extract-mediated synthesis is a simple, eco-friendly, and inexpensive method for the preparation of efficient antioxidant and photocatalytic nanoparticles. In this study, lemon peel aqueous extract was used to synthesize CuO nanoparticles (NPs) and then the obtained CuO NPs were modified using polyethylene glycol (PEG). The characteristics, optical properties, antioxidant, and photocatalytic activities of the synthesized nanoparticles were investigated. The CuO NPs and CuO/PEG NPs exhibited sphere-like morphology with an average size of 34 nm and 45 nm and optical bandgap energies of 1.2 eV and 1.5 eV, respectively. The antioxidant activity tests showed that the CuO/PEG NPs exhibited significant scavenging activity with IC50 values of 104.6 μg/mL for the β-carotene scavenging assay and 38.1 μg/mL for the ABTS scavenging assay, while CuO showed lower antioxidant activity of about 150.54 μg/mL for β-Carotene linoleic acid bleaching assay and 59.63 μg/mL for the ABTS scavenging assay. In terms of photocatalytic degradation, CuO/PEG NPs demonstrated higher activity compared to CuO NPs alone. They achieved degradation rates of 99.7% for 4-bromophenol (BP) dye and 99.5% for toluidine blue (TP) dye after 90 min, whereas CuO NPs achieved slightly lower rates. The CuO NPs and CuO/PEG NPs displayed significant photocatalytic degradation activity against amoxicillin (antibiotic), with degradation rates of 91% and 98%, respectively, after 120 min. The reaction kinetics of CuO/PEG NPs and CuO NPs followed a pseudo-first order model, with CuO/PEG NPs exhibiting a higher rate constant than CuO NPs. Overall, modifying the CuO NPs with PEG demonstrated excellent photocatalytic properties for environmental remediation and exhibited antioxidant activity, suggesting their potential use in wastewater treatment and therapeutic applications. [ABSTRACT FROM AUTHOR]

The extensive use and discharge of toluidine blue have caused serious problems to the water environment. As a green biocatalyst, laccase has the ability to decolorize the dyes, but it is limited by poor reusability and low stability. Metal–organic frameworks (MOFs) are a good platform for enzyme immobilization. However, due to the weak dispersion of MOFs, the enzyme activity is inevitably inhibited. Herein, we proposed to use graphene oxide (GO) as the dispersion medium of mesoporous ZIF-8 to construct MZIF-8/GO bi-carrier for laccase (FL) immobilization. On account of the narrower bandgap energy of FL@MZIF-8/GO (4.07 eV) than that of FL@MZIF-8 (4.69 eV), electron transport was enhanced which later increased the catalytic activity of the immobilized enzyme. Meanwhile, the improved hydrophilicity characterized by contact angle and full infiltration time further promoted the efficiency of the enzymatic reaction. Benefiting from such regulatory effects of GO, the composite showed excellent storage stability and reusability, as well as multifaceted enhancements including pH, thermal, and solvent adaptation. On the basis of the characterized synergistic effect of adsorption and degradation, FL@MZIF-8/GO was successfully applied to the degradation of toluidine blue (TB) with a removal rate of 94.8%. Even in actual treated wastewater, the highest removal rate still reached more than 80%. Based on the inner mechanism analysis and the universality study, this material is expected to be widely used in the degradation of pollutants in real water under complex environmental conditions. [ABSTRACT FROM AUTHOR]

Dye pollution is a significant environmental catastrophe, and it has so far proven to be challenging to purify dye-contaminated water effectively. Metal–organic frameworks (MOFs) have potential of removing dyes from wastewater but effectively addressing the simultaneous removal of multiple dyes remains a challenging task. In-MOF, synthesized in the round-bottom flask, sealed glass tubes, and under microwave conditions, exhibits excellent adsorption for cationic dyes. The identification and characterization of In-MOF were accomplished using several analytical techniques, including Powder X-ray diffraction, Fourier-transform infrared spectroscopy, Brunauer–Emmett–Teller analysis, Thermogravimetric analysis, Scanning electron microscope, Energy-dispersive X-ray spectroscopy, Elemental analysis, and Density functional theory for structure elucidation. In-MOF exhibits outstanding dye removal efficiency for Methylene Blue (MB: 98%; 18.6 mg/g), Azure A (AZA: 99%; 22.4 mg/g), Azure B (AZB: 98%; 17.2 mg/g), Toluidine Blue O (TOLO: 99%; 20.6 mg/g), Rhodamine B (RHB: 82%; 8.8 mg/g), and Congo red (CR: 93%; 14.2 mg/g) individually. Moreover, In-MOF demonstrates remarkable selectivity towards MB removal even in the presence of RHB. The kinetics of adsorption and reusability tests for MB indicate consistent removal capacity for the initial three cycles, but a significant decline of up to 52% is observed by the sixth cycle. These findings establish In-MOF as a promising candidate for industrial applications due to its high removal efficiency and practical synthetic approach. [ABSTRACT FROM AUTHOR]

We present the successful synthesis of a biogenic ZnO/CuO/Fe2O3 nanocomposite using an aqueous leaf extract of Ocimum Basilicum L. The confirmation of biosynthesis was achieved through UV–Visible spectrophotometry (UV–Vis), which provided evidence of ZnO/CuO/Fe2O3 NC formation. Scanning Electron Microscopy further confirmed the nanoscale size of the NC, measuring at 65 nm. X-Ray Diffraction analysis revealed a hexagonal structure for ZnO and a monoclinic structure for CuO. The successful synthesis of the environmentally friendly ZnO/CuO/Fe2O3 NC was further verified using Fourier transformed infrared (FT-IR) spectroscopy, which identified the functional groups present in the composite. Notably, the ZnO/CuO/Fe2O3 NC demonstrated exceptional degradation capabilities for toluidine blue (TB), p-toluidine (PT), and m-Toluidine (MT), with degradation rates of 99%, 99.1%, and 99.7%, respectively, within a reaction time of 120 min. The reaction kinetics followed a pseudo-first order model, with rate constant (k) values of 0.0314 min−1 and 0.0189 min−1 for TB and PT, respectively. This high rate of dye degradation can be attributed to the low band gap of the NC, which was determined to be 1.44 eV for the indirect bandgap. Furthermore, the nanocomposite exhibited excellent degradation reusability, maintaining a high degradation rate in each cycle. [ABSTRACT FROM AUTHOR]

In the present work, halloysite nanotubes modified with gold nanoparticles (AuNPs-HNT) are successfully prepared by wet chemical method for the catalytic degradation of phenothiazine dyes (azure B (AZB) and toluidine blue O (TBO)) and also cleaner reduction of 4-(4-nitrophenyl)morpholine (4NM) in the sodium borohydride (NaBH4) media. The catalyst is formulated by modifying the HNT support with a 0.964% metal loading using the HNT supports modified with 3-aminopropyl-trimethoxysilane (APTMS) coupling agent to facilitate the anchoring sites to trap the AuNPs and to prevent their agglomeration/aggregation. The AuNPs-HNT catalyst is investigated for structural and morphological characterization to get insights about the formation of the catalyst for the effective catalytic reduction of dyes and 4NM. The microscopic studies demonstrate that AuNPs (2.75 nm) are decorated on the outer surface of HNT. The as-prepared AuNPs-HNT catalyst demonstrates AZB and TBO dye degradation efficiency up to 96% in 10 and 11 min, respectively, and catalytic reduction of 4NM to 4-morpholinoaniline (MAN) is achieved up to 97% in 11 min, in the presence of NaBH4 without the formation of any by-products. The pseudo-first-order rate constant (K1) value of the AuNPs-HNT catalyst for AZB, TBO, and 4NM were calculated to be 0.0078, 0.0055, and 0.0066 s−1, respectively. Moreover, the synthesized catalyst shows an excellent reusability with stable catalytic reduction for 7 successive cycles for both the dyes and 4NM. A plausible mechanism for the catalytic dye degradation and reduction of 4NM by AuNPs-HNT catalyst is proposed as well. The obtained results clearly indicate the potential of AuNPs-HNT as an efficient catalyst for the removal of dye contaminants from the aquatic environments and cleaner reduction of 4NM to MAN, insinuating future pharmaceutical applications. [ABSTRACT FROM AUTHOR]

T4 polynucleotide kinase (T4 PNK) helps with DNA recombination and repair. In this work, a phosphate pillar[5]arene@palladium nanoparticles@reduced graphene oxide nanocomposite (PP5@PdNPs@rGO)–based electrochemical biosensor was created to identify T4 PNK activities. The PP5 used to complex toluidine blue (TB) guest molecules is water-soluble. With T4 PNK and ATP, the substrate DNA, which included a 5ʹ-hydroxyl group, initially self-assembled over the gold electrode surface by chemical adsorption of the thiol units. Strong phosphate-Zr4+-phosphate chemistry allowed Zr4+ to act as a bridge between phosphorylated DNA and PP5@PdNPs@rGO. Through a supramolecular host–guest recognition connection, TB molecules were able to penetrate the PP5 cavity, where they produced a stronger electrochemical response. With a 5 × 10−7 U mL−1 detection limit, the electrochemical signal is linear in the 10−6 to 1 U mL−1 T4 PNK concentration range. It was also effective in measuring HeLa cell lysate–related PNK activities and screening PNK inhibitors. Nucleotide kinase–target drug development, clinical diagnostics, and screening for inhibitors all stand to benefit greatly from the suggested technology, which offers a unique sensing mechanism for kinase activity measurement. [ABSTRACT FROM AUTHOR]

A poly(toluidine blue) modified glassy carbon electrode (PTB/GCE) was prepared by two-step electropolymerization, and was used for the stripping voltammetric analysis of trace uranium for the first time. The results showed that the PTB modified electrode had higher sensitivity of uranium detection compared with the glassy carbon electrode, and the standard addition method was satisfactory for seawater, river water and tap water. The PTB/GCE can be used as a mercury-free electrode for the electrochemical detection of uranyl ions in the environmental water, which provides a new way for the rapid detection of trace uranium. [ABSTRACT FROM AUTHOR]

Background: Screening and early diagnosis of oral squamous cell carcinoma (OSCC) are directly associated with increased survival rate and improved prognosis. Noninvasive diagnostic tools have been implemented in the early detection as toluidine blue staining, optical imaging, and oral cytology. This study aimed to assess and compare the presence of micronuclei (MN) in oral exfoliative cytology of healthy controls, subjects exposed to high-risk factors for oral cancer, subjects with oral potentially malignant lesions (OPMLs), and those with malignant oral lesions. Subjects and methods: A total number of 92 subjects were divided into 46 healthy controls with no oral mucosal lesions (23 with no evidence of cancer risk factors and 23 with cancer risk factors), 23 with OPMLs and 23 with oral malignant lesions. All the 92 participants were subjected to cytological sampling for detection of MN. The final diagnosis of the oral lesions was confirmed by the histopathological picture and compared to the cytological results. Results: The results showed that the diagnostic accuracy of MN was higher in OPMLs group (95.2%). The sensitivity of MN test in malignant group was much lower (52.2%); however, all the cytological criteria of malignancy were markedly detected as compared to the OPMLs group. Conclusions: Conventional oral cytology supported by MN is highly beneficial as adjunctive tool in the screening for early detection of dysplastic oral lesions. [ABSTRACT FROM AUTHOR]

Structural and functional parameters of mast cells (MC) in rat mammary glands during experimentally provoked breast cancer were studied by the cytogram, index of cell saturation with secretory products, and degranulation index. The cytogram was calculated in histological sections stained with toluidine blue. The functional conditions of MC were determined by electron microscopy. The study revealed expansion of MC population and activation of their functional state evidenced by significant prevalence of cells with high degranulation degree and reduced saturation index. At the ultrastructural level, MC were characterized by deformation, polymorphism, paleness of secretory granules, and elevation of the number of lacunae in the cytoplasm. The study showed that MC are active players in tumor microenvironment. Remembering heteromorphism of MC, further study of their role in pathogenesis of various tumor diseases seems promising. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the influence of IR (λ850 ± 10 nm) and violet (λ405 ± 10 nm) LED phototherapy on total mast cells counts and its ability to influence mast cell degranulation. For this, 27 Wistar rats were used and were randomly distributed into three groups: control, IR LED, and violet LED. When indicated, irradiation done and they were sacrificed, had their tongue removed immediately, 20-min, 45-min, and 2-h after irradiation. Samples were processed to wax, cut, and stained with Toluidine Blue. Intact and degranulated mast cells were counted under light microscopy, and statistical analysis was carried out. In the superficial connective tissue and muscular tissues, violet LED light caused a significant increase in both total number and degranulated mast cells when compared to the control group immediately after irradiation. The degranulation indexes were higher in the groups irradiated with Violet light, both in superficial connective tissue and muscular tissues in relation to the timing. Irradiation with IR LED caused immediate increase in the total number and degranulated of mast cells when compared to the control group only in the superficial connective tissue. In all times observed, the highest total amount of mast cells was seen immediately after irradiation, except in the muscular tissue, which presented the highest amount after 20-min. It was concluded that IR and violet LED light were able to increase the number of mast cells and inducing degranulation in oral mucosa. However, considering that violet LED light can be harmful in periodontal disease, it seems that the use of IR LED light could be the best option in Dentistry. [ABSTRACT FROM AUTHOR]

In this paper, a new multifunctional nanocomposite was synthesized to combine both the advantages of sorption properties and antibacterial efficiency. The thiourea polyurethane/zinc oxide nanocomposite was prepared by coupling thiourea polyurethane foam and zinc oxide nanoparticles. The thiourea polyurethane/zinc oxide was characterized using several tools. The change in the surface chemistry of the thiourea polyurethane/zinc oxide nanocomposite (surface area was increased from 2.8 to 10.6 m2/g), due to the attached functional groups, increased its sorption properties. Also, the thiourea polyurethane/zinc oxide showed a crystalline phase, embedded inside the amorphous structure, with good thermal stability to 230 °C. The pore volume and radius of thiourea polyurethane/zinc oxide were 0.017 cm3/g and 3.4 nm with mesopores/micropores ratio of 11:1. It combines collecting pollutants on its surface, the ability to degrade these pollutants, and a good antibacterial effect. Thiourea polyurethane/zinc oxide was applied for the removing toluidine blue and brilliant green basic dyes from the wastewater. Several parameters such as pH, dye concentration, temperature, and time were studied. The removal percentages of brilliant green and toluidine blue dyes from the wastewater were 92.4–98.2% at pH 3–12. Also, the removing rate of the dyes was very fast (3–5 min) at 25 °C. The estimated capacities of new nanocomposite for toluidine blue and brilliant green were 92 and 227 mg/g. The removal process was spontaneous (ΔG was − 7.3 and − 8.6). As a result, the applicability of thiourea polyurethane/zinc oxide as an effective renewable antibacterial sorbent for removing cationic dyes was maintained. [ABSTRACT FROM AUTHOR]

Human epididymis protein 4 (HE4) is a highly sensitive and specific biomarker for diagnosing ovarian cancer. In this work, a new electrochemical aptasensor for rapid and sensitive detection of HE4 was constructed. Firstly, metal–organic framework/ketjen black (NH2-MIL-53(Al)/KB) composite with abundant amino groups and large specific surface area was prepared as signal amplifier to anchor gold nanoparticles (AuNPs). Subsequently, signal probe (SP) and a large amount of electroactive toluidine blue (Tb) were attached on AuNPs to form the tracer label, producing a distinct electrochemical detection signal. Moreover, Ni3(HITP)2 was modified by poly(diallyldimethylammonium chloride, PDDA) to obtain PDDA/Ni3(HITP)2, and then it was covered with deposited gold nanocrystals (DpAu) to form the sensing platform, which increased the electrical conductivity, specific surface area and the loading of triple-helix assembled probes (TAP). Target HE4 was bound to its aptamer in the outer layer of the TAP, leaving a separate capture probe (CP) on the electrode surface that can bind with the tracer label. The proposed aptasensor for HE4 detection showed a wide linear range from 1 to 10 nM with a detection limit of 0.41 fM. In addition, the aptasensor has good selectivity, stability and reproducibility. More importantly, this method showed satisfactory results in clinical serum sample analysis compared with ELISA, possessing application prospects in ovarian cancer diagnosis. [ABSTRACT FROM AUTHOR]

A semi-interpenetrating network consisting of karaya gum-graft-polyacrylamide and poly(vinyl alcohol) has been developed as an adsorbent material for dye removal applications. The system was made by microwave irradiation of the aqueous mixture of karaya gum, acrylamide, N,N′-methylenebisacrylamide, and poly (vinyl alcohol); and was characterized by FTIR, TGA, SEM, and XRD techniques. It was evaluated as an adsorbent material for removal of ionic dyes from aqueous solutions. Appreciably high adsorption capacity is exhibited by the material towards cationic dyes as indicated by values of maximum adsorption capacity of 82.28, 72.94, 48.75 and 34.67 mg/g towards adsorption of cationic dyes, namely methylene blue (MB), crystal violet (CV), rhodamine B (Rh B), and toluidine blue (TB). Relatively low value of 21.66 mg/g is obtained for anionic dye indigo carmine (IC). The kinetic data of dye adsorption best fitted with the pseudo-first-order kinetic model with R2 values of 0.979, 0.989, 0.946, 0.991, and 0.986 for TB, MB, CV, Rh B and IC, respectively. The isotherm data fit well with the Freundlich model with R2 values of 0.986, 0.955, 0.990, 0.980 and 0.993 for TB, MB, CV, Rh B, and IC, respectively, indicating the heterogeneous and multilayer adsorption. The thermodynamic studies showed dye adsorption to be an exothermic and spontaneous process. [ABSTRACT FROM AUTHOR]

Toluidine blue O (TBO) is a phenothiazine dye that, due to its photochemical characteristics and high affinity for biomembranes, has been revealed as a new photosensitizer (PS) option for antimicrobial photodynamic therapy (PDT). This points to a possible association with membranous organelles like mitochondrion. Therefore, here we investigated its effects on mitochondrial bioenergetic functions both in the dark and under photostimulation. Two experimental systems were utilized: (a) isolated rat liver mitochondria and (b) isolated perfused rat liver. Our data revealed that, independently of photostimulation, TBO presented affinity for mitochondria. Under photostimulation, TBO increased the protein carbonylation and lipid peroxidation levels (up to 109.40 and 119.87%, respectively) and decreased the reduced glutathione levels (59.72%) in mitochondria. TBO also uncoupled oxidative phosphorylation and photoinactivated the respiratory chain complexes I, II, and IV, as well as the FoF1-ATP synthase complex. Without photostimulation, TBO caused uncoupling of oxidative phosphorylation and loss of inner mitochondrial membrane integrity and inhibited very strongly succinate oxidase activity. TBO's uncoupling effect was clearly seen in intact livers where it stimulated oxygen consumption at concentrations of 20 and 40 μM. Additionally, TBO (40 μM) reduced cellular ATP levels (52.46%) and ATP/ADP (45.98%) and ATP/AMP (74.17%) ratios. Consequently, TBO inhibited gluconeogenesis and ureagenesis whereas it stimulated glycogenolysis and glycolysis. In conclusion, we have revealed for the first time that the efficiency of TBO as a PS may be linked to its ability to photodynamically inhibit oxidative phosphorylation. In contrast, TBO is harmful to mitochondrial energy metabolism even without photostimulation, which may lead to adverse effects when used in PDT. [ABSTRACT FROM AUTHOR]

Calcium phosphate forms particles under excessive urinary excretion of phosphate in the kidney. While the formation of calcium phosphate particles (CaPs) has been implicated in the damage to renal tubular cells and renal dysfunction, clarifying the ultrastructural information and the elemental composition of the small CaPs in the wide areas of kidney tissue has been technically difficult. This study introduces correlative and sequential light as well as electron microscopic CaP observation in the kidney tissue by combining fluorescent staining for CaPs and energy-dispersive X-ray spectroscopy (EDS) in scanning electron microscopy (SEM) on resin sections prepared using high-pressure freezing and freeze substitution. CaPs formed in mouse kidneys under long-term feeding of a high-phosphate diet were clearly visualized on resin sections by fluorescence-conjugated alendronate derivatives and toluidine blue metachromasia. These CaPs were verified by correlative observation with EDS. Furthermore, small CaPs formed in the kidney under short-term feeding were detected using fluorescent probes. The elemental composition of the particles, including calcium and magnesium, was identified following EDS analyses. These results suggest that the correlative microscopy approach is helpful for observing in situ distribution and elemental composition of CaPs in the kidney and contributing to studies regarding CaP formation-associated pathophysiology. [ABSTRACT FROM AUTHOR]

The surface sulfate groups of cotton-derived cellulose nanowhiskers (CNWs) were quantified using three different methods: conductometric titration, adsorption of toluidine blue O (TBO), and sulfur elemental analysis. The former two methods indicated comparable values, whereas the last method indicated a considerable overestimation of up to 1.4 times the sulfate content compared to the values obtained by titration. This overestimation may be due to the inaccessibility of the sulfates inside the CNWs to the titrant and TBO, and/or the presence of non-sulfate sulfur elements. In addition to the quantitativeness of the TBO adsorption method, the present results also demonstrate the desulfation efficiency under several different desulfation conditions, where the sulfate content decreased by only half after alkali-mediated desulfation even with 2 M NaOH, whereas quite effective desulfation (ca. 90% decrease) was achieved with dilute HCl (0.25 M). [ABSTRACT FROM AUTHOR]

The relevance of the work is related to the need to develop methods for determining individual opportunities to minimize the harmful effects of ionizing radiation. Experiments were conducted to search for informative indicators of the structure of mast cells to predict the individual effectiveness of the radioprotective effect of hypoxia. A study was performed on white male rats. In 81 animals, before the main experiment, a hypoxic sample was carried out once, by placing the rats in a gas mixture containing 8% oxygen and 92% nitrogen (GGS-8). In each of these rats, a biopsy of the skin of the auricle was performed immediately before the test and at the time of its completion. In the biopsies, the content of various forms of mast cells detected by the method of metachromasia with toluidine blue was determined. After a week, all animals of the main group (81 rats) were exposed to total γ-irradiation at a dose of 11 Gy under the protection of GGS-8. Rats in the control group (63 rats) were irradiated in the same dose in the air. The life expectancy after the exposure was assessed. The radioprotective effect of the gas hypoxic mixture was confirmed, which is expressed in an increase in the average life expectancy of irradiated rats by 2.2 times. A weak relationship between individual quantitative characteristics of mast cells and the life expectancy of irradiated animals is shown. A formal expression of the equation of regression of the number of mast cells on the life expectancy of animals irradiated under the protection of hypoxia has been created. On the basis of regression analysis, groups of animals in which the highest radioprotective effect of hypoxia was manifested were revealed. The minimum life expectancy after irradiation is characteristic of rats with extreme values of the mast cell reactivity index, and all surviving cases are noted in rats with average values of the indicator. Among them there was not a single animal with an intestinal form of death. This indicates the possibility of predicting the radioprotective effect of hypoxia based on an assessment of the functional activity of mast cells. [ABSTRACT FROM AUTHOR]

Early detection is a major step in the success of cancer therapy. Histopathology report is considered as the gold standard in the formulation of management protocol of any malignancy worldwide. But unfortunately, there is a delay in the detection of oral cancer very often due to inconclusive histopathology reports. The main reason behind it is obtaining a biopsy specimen from the non-representative area of the lesion. A hospital-based evaluation of the role of Toluidine Blue dye, used as an adjunctive method prior to biopsy was conducted in a tertiary care hospital on 200 patients presenting with oral lesions persistent for more than 3 weeks. The participants were divided into two equal groups by alternate sampling. In one group biopsy was taken by clinical judgment and in others, Toluidine Blue was used prior to obtaining a biopsy to decide the area to be biopsied. Data was collected using a predesigned proforma and was analyzed with the help of SPSS version 20. Results in two groups were compared with respect to sensitivity, specificity, positive and negative predictive values, false positive and false negative percentages. The Sensitivity, Specificity, Positive Predictive Value and Negative Predictive Value of wedge biopsy without staining were 73.68, 58.14, 70.00, and 62.50% respectively. These values were 95.08, 82.05, 89.23, and 91.43% respectively when Toluidine Blue staining was done as an adjunctive before the biopsy procedure. These results indicate the promising role of Toluidine blue staining before the biopsy to diagnose oral malignancy more efficiently than obtaining biopsy specimens on clinical assessment only and in avoiding the delay in initiating the treatment in case of oral malignant lesions. [ABSTRACT FROM AUTHOR]

Rationale: Major depression has been an area of extensive research during the last decades, for it represents a leading cause of disability and suicide. The stark rise of depression rates influenced by life stressors, economic threats, pandemic era, and resistance to classical treatments, has made the disorder rather challenging. Adult hippocampal neurogenesis and plasticity are particularly sensitive to the dynamic interplay between autophagy and inflammation. In fact, the intricate balance between the two processes contributes to neuronal homeostasis and survival.Having demonstrated promising potentials in AMPK activation, a major metabolic sensor and autophagy regulator, empagliflozin (Empa) was investigated for possible antidepressant properties in the reserpine rat model of depression.While the reserpine protocol elicited behavioral, biochemical, and histopathological changes relevant to depression, Empa outstandingly hindered these pathological perturbations. Importantly, hippocampal autophagic response markedly declined with reserpine which disrupted the AMPK/mTOR/Beclin1/LC3B machinery and, conversely, neuro-inflammation prevailed under the influence of the NLRP3 inflammasome together with oxidative/nitrative stress. Consequently, AMPK-mediated neurotrophins secretion obviously deteriorated through PKCζ/NF-κB/BDNF/CREB signal restriction. Empa restored hippocampal monoamines and autophagy/inflammation balance, driven by AMPK activation. By promoting the atypical PKCζ phosphorylation (Thr403) which subsequently phosphorylates NF-κB at Ser311, AMPK successfully reinforced BDNF/CREB signal and hippocampal neuroplasticity. The latter finding was supported by hippocampal CA3 toluidine blue staining to reveal intact neurons.The current study highlights an interesting role for Empa as a regulator of autophagic and inflammatory responses in the pathology of depression. The study also pinpoints an unusual contribution for NF-κB in neurotrophins secretion via AMPK/PKCζ/NF-κB/BDNF/CREB signal transduction. Accordingly, Empa can have special benefits in diabetic patients with depressive symptoms.The influence of p-NF-κB (Ser311) on NLRP3 inflammasome assembly and activation has not been investigated, which can represent an interesting point for further research.Objectives: Major depression has been an area of extensive research during the last decades, for it represents a leading cause of disability and suicide. The stark rise of depression rates influenced by life stressors, economic threats, pandemic era, and resistance to classical treatments, has made the disorder rather challenging. Adult hippocampal neurogenesis and plasticity are particularly sensitive to the dynamic interplay between autophagy and inflammation. In fact, the intricate balance between the two processes contributes to neuronal homeostasis and survival.Having demonstrated promising potentials in AMPK activation, a major metabolic sensor and autophagy regulator, empagliflozin (Empa) was investigated for possible antidepressant properties in the reserpine rat model of depression.While the reserpine protocol elicited behavioral, biochemical, and histopathological changes relevant to depression, Empa outstandingly hindered these pathological perturbations. Importantly, hippocampal autophagic response markedly declined with reserpine which disrupted the AMPK/mTOR/Beclin1/LC3B machinery and, conversely, neuro-inflammation prevailed under the influence of the NLRP3 inflammasome together with oxidative/nitrative stress. Consequently, AMPK-mediated neurotrophins secretion obviously deteriorated through PKCζ/NF-κB/BDNF/CREB signal restriction. Empa restored hippocampal monoamines and autophagy/inflammation balance, driven by AMPK activation. By promoting the atypical PKCζ phosphorylation (Thr403) which subsequently phosphorylates NF-κB at Ser311, AMPK successfully reinforced BDNF/CREB signal and hippocampal neuroplasticity. The latter finding was supported by hippocampal CA3 toluidine blue staining to reveal intact neurons.The current study highlights an interesting role for Empa as a regulator of autophagic and inflammatory responses in the pathology of depression. The study also pinpoints an unusual contribution for NF-κB in neurotrophins secretion via AMPK/PKCζ/NF-κB/BDNF/CREB signal transduction. Accordingly, Empa can have special benefits in diabetic patients with depressive symptoms.The influence of p-NF-κB (Ser311) on NLRP3 inflammasome assembly and activation has not been investigated, which can represent an interesting point for further research.Results: Major depression has been an area of extensive research during the last decades, for it represents a leading cause of disability and suicide. The stark rise of depression rates influenced by life stressors, economic threats, pandemic era, and resistance to classical treatments, has made the disorder rather challenging. Adult hippocampal neurogenesis and plasticity are particularly sensitive to the dynamic interplay between autophagy and inflammation. In fact, the intricate balance between the two processes contributes to neuronal homeostasis and survival.Having demonstrated promising potentials in AMPK activation, a major metabolic sensor and autophagy regulator, empagliflozin (Empa) was investigated for possible antidepressant properties in the reserpine rat model of depression.While the reserpine protocol elicited behavioral, biochemical, and histopathological changes relevant to depression, Empa outstandingly hindered these pathological perturbations. Importantly, hippocampal autophagic response markedly declined with reserpine which disrupted the AMPK/mTOR/Beclin1/LC3B machinery and, conversely, neuro-inflammation prevailed under the influence of the NLRP3 inflammasome together with oxidative/nitrative stress. Consequently, AMPK-mediated neurotrophins secretion obviously deteriorated through PKCζ/NF-κB/BDNF/CREB signal restriction. Empa restored hippocampal monoamines and autophagy/inflammation balance, driven by AMPK activation. By promoting the atypical PKCζ phosphorylation (Thr403) which subsequently phosphorylates NF-κB at Ser311, AMPK successfully reinforced BDNF/CREB signal and hippocampal neuroplasticity. The latter finding was supported by hippocampal CA3 toluidine blue staining to reveal intact neurons.The current study highlights an interesting role for Empa as a regulator of autophagic and inflammatory responses in the pathology of depression. The study also pinpoints an unusual contribution for NF-κB in neurotrophins secretion via AMPK/PKCζ/NF-κB/BDNF/CREB signal transduction. Accordingly, Empa can have special benefits in diabetic patients with depressive symptoms.The influence of p-NF-κB (Ser311) on NLRP3 inflammasome assembly and activation has not been investigated, which can represent an interesting point for further research.Conclusion: Major depression has been an area of extensive research during the last decades, for it represents a leading cause of disability and suicide. The stark rise of depression rates influenced by life stressors, economic threats, pandemic era, and resistance to classical treatments, has made the disorder rather challenging. Adult hippocampal neurogenesis and plasticity are particularly sensitive to the dynamic interplay between autophagy and inflammation. In fact, the intricate balance between the two processes contributes to neuronal homeostasis and survival.Having demonstrated promising potentials in AMPK activation, a major metabolic sensor and autophagy regulator, empagliflozin (Empa) was investigated for possible antidepressant properties in the reserpine rat model of depression.While the reserpine protocol elicited behavioral, biochemical, and histopathological changes relevant to depression, Empa outstandingly hindered these pathological perturbations. Importantly, hippocampal autophagic response markedly declined with reserpine which disrupted the AMPK/mTOR/Beclin1/LC3B machinery and, conversely, neuro-inflammation prevailed under the influence of the NLRP3 inflammasome together with oxidative/nitrative stress. Consequently, AMPK-mediated neurotrophins secretion obviously deteriorated through PKCζ/NF-κB/BDNF/CREB signal restriction. Empa restored hippocampal monoamines and autophagy/inflammation balance, driven by AMPK activation. By promoting the atypical PKCζ phosphorylation (Thr403) which subsequently phosphorylates NF-κB at Ser311, AMPK successfully reinforced BDNF/CREB signal and hippocampal neuroplasticity. The latter finding was supported by hippocampal CA3 toluidine blue staining to reveal intact neurons.The current study highlights an interesting role for Empa as a regulator of autophagic and inflammatory responses in the pathology of depression. The study also pinpoints an unusual contribution for NF-κB in neurotrophins secretion via AMPK/PKCζ/NF-κB/BDNF/CREB signal transduction. Accordingly, Empa can have special benefits in diabetic patients with depressive symptoms.The influence of p-NF-κB (Ser311) on NLRP3 inflammasome assembly and activation has not been investigated, which can represent an interesting point for further research.Limitations: Major depression has been an area of extensive research during the last decades, for it represents a leading cause of disability and suicide. The stark rise of depression rates influenced by life stressors, economic threats, pandemic era, and resistance to classical treatments, has made the disorder rather challenging. Adult hippocampal neurogenesis and plasticity are particularly sensitive to the dynamic interplay between autophagy and inflammation. In fact, the intricate balance between the two processes contributes to neuronal homeostasis and survival.Having demonstrated promising potentials in AMPK activation, a major metabolic sensor and autophagy regulator, empagliflozin (Empa) was investigated for possible antidepressant properties in the reserpine rat model of depression.While the reserpine protocol elicited behavioral, biochemical, and histopathological changes relevant to depression, Empa outstandingly hindered these pathological perturbations. Importantly, hippocampal autophagic response markedly declined with reserpine which disrupted the AMPK/mTOR/Beclin1/LC3B machinery and, conversely, neuro-inflammation prevailed under the influence of the NLRP3 inflammasome together with oxidative/nitrative stress. Consequently, AMPK-mediated neurotrophins secretion obviously deteriorated through PKCζ/NF-κB/BDNF/CREB signal restriction. Empa restored hippocampal monoamines and autophagy/inflammation balance, driven by AMPK activation. By promoting the atypical PKCζ phosphorylation (Thr403) which subsequently phosphorylates NF-κB at Ser311, AMPK successfully reinforced BDNF/CREB signal and hippocampal neuroplasticity. The latter finding was supported by hippocampal CA3 toluidine blue staining to reveal intact neurons.The current study highlights an interesting role for Empa as a regulator of autophagic and inflammatory responses in the pathology of depression. The study also pinpoints an unusual contribution for NF-κB in neurotrophins secretion via AMPK/PKCζ/NF-κB/BDNF/CREB signal transduction. Accordingly, Empa can have special benefits in diabetic patients with depressive symptoms.The influence of p-NF-κB (Ser311) on NLRP3 inflammasome assembly and activation has not been investigated, which can represent an interesting point for further research.Graphical abstract: Major depression has been an area of extensive research during the last decades, for it represents a leading cause of disability and suicide. The stark rise of depression rates influenced by life stressors, economic threats, pandemic era, and resistance to classical treatments, has made the disorder rather challenging. Adult hippocampal neurogenesis and plasticity are particularly sensitive to the dynamic interplay between autophagy and inflammation. In fact, the intricate balance between the two processes contributes to neuronal homeostasis and survival.Having demonstrated promising potentials in AMPK activation, a major metabolic sensor and autophagy regulator, empagliflozin (Empa) was investigated for possible antidepressant properties in the reserpine rat model of depression.While the reserpine protocol elicited behavioral, biochemical, and histopathological changes relevant to depression, Empa outstandingly hindered these pathological perturbations. Importantly, hippocampal autophagic response markedly declined with reserpine which disrupted the AMPK/mTOR/Beclin1/LC3B machinery and, conversely, neuro-inflammation prevailed under the influence of the NLRP3 inflammasome together with oxidative/nitrative stress. Consequently, AMPK-mediated neurotrophins secretion obviously deteriorated through PKCζ/NF-κB/BDNF/CREB signal restriction. Empa restored hippocampal monoamines and autophagy/inflammation balance, driven by AMPK activation. By promoting the atypical PKCζ phosphorylation (Thr403) which subsequently phosphorylates NF-κB at Ser311, AMPK successfully reinforced BDNF/CREB signal and hippocampal neuroplasticity. The latter finding was supported by hippocampal CA3 toluidine blue staining to reveal intact neurons.The current study highlights an interesting role for Empa as a regulator of autophagic and inflammatory responses in the pathology of depression. The study also pinpoints an unusual contribution for NF-κB in neurotrophins secretion via AMPK/PKCζ/NF-κB/BDNF/CREB signal transduction. Accordingly, Empa can have special benefits in diabetic patients with depressive symptoms.The influence of p-NF-κB (Ser311) on NLRP3 inflammasome assembly and activation has not been investigated, which can represent an interesting point for further research. [ABSTRACT FROM AUTHOR]

Introduction: Articular cartilage is the major affected tissue during the development of osteoarthritis (OA) in temporomandibular joint (TMJ). The core circadian rhythm molecule Bmal1 regulates chondrocyte proliferation, differentiation and apoptosis; however, its roles in condylar cartilage function and in TMJ OA have not been fully elucidated.TMJ OA mouse model was induced by unilateral anterior crossbite (UAC) and Bmal1 protein expression in condylar cartilage were examined by western blot analysis. To determine the role of Bmal1 in TMJ OA, we generated cartilage-specific Bmal1 conditional knockout (cKO) mice (Bmal1Agc1CreER mice) and hematoxylin and eosin staining, toluidine blue and Safranin O/fast green, immunohistochemistry, TUNEL assay, real-time PCR analysis and Western blot assay were followed.Bmal1 expression was reduced in condylar cartilage in a TMJ OA mouse model induced by UAC. The Bmal1 cKO mice displayed decreased cartilage matrix synthesis, reduced chondrocyte proliferation, increased chondrocyte hypertrophy and apoptosis as well as the upregulation of YAP expression in TMJ condylar cartilage.We demonstrated that Bmal1 was essential for TMJ tissue homeostasis and loss-of-function of Bmal1 in chondrocytes leads to the development of TMJ OA.Materials and methods: Articular cartilage is the major affected tissue during the development of osteoarthritis (OA) in temporomandibular joint (TMJ). The core circadian rhythm molecule Bmal1 regulates chondrocyte proliferation, differentiation and apoptosis; however, its roles in condylar cartilage function and in TMJ OA have not been fully elucidated.TMJ OA mouse model was induced by unilateral anterior crossbite (UAC) and Bmal1 protein expression in condylar cartilage were examined by western blot analysis. To determine the role of Bmal1 in TMJ OA, we generated cartilage-specific Bmal1 conditional knockout (cKO) mice (Bmal1Agc1CreER mice) and hematoxylin and eosin staining, toluidine blue and Safranin O/fast green, immunohistochemistry, TUNEL assay, real-time PCR analysis and Western blot assay were followed.Bmal1 expression was reduced in condylar cartilage in a TMJ OA mouse model induced by UAC. The Bmal1 cKO mice displayed decreased cartilage matrix synthesis, reduced chondrocyte proliferation, increased chondrocyte hypertrophy and apoptosis as well as the upregulation of YAP expression in TMJ condylar cartilage.We demonstrated that Bmal1 was essential for TMJ tissue homeostasis and loss-of-function of Bmal1 in chondrocytes leads to the development of TMJ OA.Results: Articular cartilage is the major affected tissue during the development of osteoarthritis (OA) in temporomandibular joint (TMJ). The core circadian rhythm molecule Bmal1 regulates chondrocyte proliferation, differentiation and apoptosis; however, its roles in condylar cartilage function and in TMJ OA have not been fully elucidated.TMJ OA mouse model was induced by unilateral anterior crossbite (UAC) and Bmal1 protein expression in condylar cartilage were examined by western blot analysis. To determine the role of Bmal1 in TMJ OA, we generated cartilage-specific Bmal1 conditional knockout (cKO) mice (Bmal1Agc1CreER mice) and hematoxylin and eosin staining, toluidine blue and Safranin O/fast green, immunohistochemistry, TUNEL assay, real-time PCR analysis and Western blot assay were followed.Bmal1 expression was reduced in condylar cartilage in a TMJ OA mouse model induced by UAC. The Bmal1 cKO mice displayed decreased cartilage matrix synthesis, reduced chondrocyte proliferation, increased chondrocyte hypertrophy and apoptosis as well as the upregulation of YAP expression in TMJ condylar cartilage.We demonstrated that Bmal1 was essential for TMJ tissue homeostasis and loss-of-function of Bmal1 in chondrocytes leads to the development of TMJ OA.Conclusions: Articular cartilage is the major affected tissue during the development of osteoarthritis (OA) in temporomandibular joint (TMJ). The core circadian rhythm molecule Bmal1 regulates chondrocyte proliferation, differentiation and apoptosis; however, its roles in condylar cartilage function and in TMJ OA have not been fully elucidated.TMJ OA mouse model was induced by unilateral anterior crossbite (UAC) and Bmal1 protein expression in condylar cartilage were examined by western blot analysis. To determine the role of Bmal1 in TMJ OA, we generated cartilage-specific Bmal1 conditional knockout (cKO) mice (Bmal1Agc1CreER mice) and hematoxylin and eosin staining, toluidine blue and Safranin O/fast green, immunohistochemistry, TUNEL assay, real-time PCR analysis and Western blot assay were followed.Bmal1 expression was reduced in condylar cartilage in a TMJ OA mouse model induced by UAC. The Bmal1 cKO mice displayed decreased cartilage matrix synthesis, reduced chondrocyte proliferation, increased chondrocyte hypertrophy and apoptosis as well as the upregulation of YAP expression in TMJ condylar cartilage.We demonstrated that Bmal1 was essential for TMJ tissue homeostasis and loss-of-function of Bmal1 in chondrocytes leads to the development of TMJ OA. [ABSTRACT FROM AUTHOR]

Background: Cypripedium subtropicum is a unique, endangered lady's slipper orchid with evergreen leaves on non-dormant shoots that is native to southwestern China. This study documents the major developmental events in C. subtropicum seed development from fertilization to seed maturity, determines the optimum period for seed collection, and examines the cytokinin requirements for asymbiotic germination and protocorm survival. Results: Structural studies revealed that embryo development proceeded after successful fertilization at 60 days after pollination (DAP). At 105 DAP, a globular embryo with the shrinking inner seed coat was observed, and seeds collected at this time point exhibited optimal germination. After 120 DAP, most seeds had a mature embryo within the capsule, and within the cells of the embryo proper, numerous proteins/lipid bodies were present as the main storage products. In addition, the inner seed coat had compressed into a thin layer that tightly enclosed the embryo, while the outer seed coat had progressively elongated, resulting in a hair-like appearance of the mature seed. Histochemical staining using Nile red and toluidine blue O (TBO) indicated that the lignified inner and outer seed coats may lead to coat-imposed dormancy. Seeds collected at this stage germinated poorly. Analyses of cytokinin preferences and optimal concentrations for germination and protocorm survival showed that both 6-(γ,γ-dimethylallylamino) purine (2iP) and 6-benzylaminopurine (BA) enhanced germination compared with the control, although higher concentrations of BA (4 and 8 μM) suppressed germination. The protocorm survival rate improved with increasing 2iP concentration. Conclusions: This study provides a reproducible procedure for culturing immature seeds of C. subtropicum based on a defined time schedule of seed development. In addition, the cytokinin 2iP was shown to improve germination and protocorm survival. This study provides a scientific basis for seedling establishment through asymbiotic seed culture for further reintroduction efforts. [ABSTRACT FROM AUTHOR]

Introduction: Chondroprogenitors (CPCs) have emerged as a promising cellular therapy for cartilage-related pathologies due to their inherent primed chondrogenic potential. Studies report that the addition of growth factors such as parathyroid hormone (PTH) and Bone Morphogenic Protein (BMP) enhance the chondroinducive potential in chondrocytes and mesenchymal stem cells. This study evaluated if supplementation of the standard culture medium for cell expansion with 1–34 PTH and BMP-9 would enhance the chondrogenic potential of CPCs and reduce their hypertrophic tendency. Methods: Human chondrocytes were isolated from patients undergoing total knee replacement for osteoarthritis (n = 3). Following fibronectin adhesion assay, passage 1 CPCs were divided and further expanded under three culture conditions (a) control, i.e., cells continued under standard culture conditions, (b) 1–34 PTH group, additional intermittent 6 h exposure with 1–34 PTH and (c) BMP-9 group, additional BMP-9 during culture expansion. All the groups were evaluated for population-doubling, cell cycle analysis, surface marker and gene expression for chondrogenesis, hypertrophy, multilineage differentiation and GAG (glycosaminoglycan)/DNA following chondrogenic differentiation. Results: Concerning growth kinetics, the BMP-9 group exhibited a significantly lower S-phase and population-doubling when compared to the other two groups. Qualitative analysis for chondrogenic potential (Alcian blue, Safranin O staining and Toluidine blue for GAG) revealed that the BMP-9 group exhibited the highest uptake. The BMP-9 group also showed significantly higher COL2A1 expression than the control group, with no change in the hypertrophy marker expression. Conclusion: BMP-9 can potentially be used as an additive for CPCs expansion, to enhance their chondrogenic potential without affecting their low hypertrophic tendency. The mitigating effects of 1-34PTH on hypertrophy would benefit further investigation when used in combination with BMP-9 to enhance chondrogenesis whilst reducing hypertrophy. [ABSTRACT FROM AUTHOR]

Efficient transport of solutes in the cerebrospinal fluid (CSF) plays a critical role in their clearance from the brain. Convective bulk flow of solutes in the CSF in the perivascular space (PVS) is considered one of the important mechanisms behind solute movement in the brain, before their ultimate drainage to the systemic lymphatic system. Acoustic pressure waves can impose radiation force on a medium in its path, inducing localized and directional fluidic flow, known as acoustic streaming. We transcranially applied low-intensity focused ultrasound (FUS) to rats that received an intracisternal injection of fluorescent CSF tracers (dextran and ovalbumin, having two different molecular weights–Mw). The sonication pulsing parameter was determined on the set that propelled the aqueous solution of toluidine blue O dye into a porous media (melamine foam) at the highest level of infiltration. Fluorescence imaging of the brain showed that application of FUS increased the uptake of ovalbumin at the sonicated plane, particularly around the ventricles, whereas the uptake of high-Mw dextran was unaffected. Numerical simulation showed that the effects of sonication were non-thermal. Sonication did not alter the animals' behavior or disrupt the blood-brain barrier (BBB) while yielding normal brain histology. The results suggest that FUS may serve as a new non-invasive means to promote interstitial CSF solute transport in a region-specific manner without disrupting the BBB, providing potential for enhanced clearance of waste products from the brain. [ABSTRACT FROM AUTHOR]

Objectives: The aim of this study is to evaluate the influence of the antimicrobial photodynamic therapy with phycocyanin and toluidine blue on the bond strength of resin composite to the sound dentin. Materials and methods: The samples include 120 tooth blocks taken from 60 human extracted sound third molar teeth. They were randomly divided into 12 groups and disinfected with two different photosensitizers (Phycocyanin and Toluidine blue) activated by 635 nm diode laser for aPDT procedure. Then two different protocols (total-etch and self-etch) of universal adhesive system (G-Premio BOND, GC Dental Products CORP. Japan) were applied. The samples then filled with resin composite (Gradia® Direct, GC Dental Products CORP. Japan). After 10000 cycles of thermocycling, all samples were subjected to the micro shear bond strength (μSBS) test using a universal testing machine (1 mm/min). The data were analyzed with One Way ANOVA. Results: The effect of aPDT using both photosensitizers (phycocyanin and toluidine blue O) on the bonding strength to dentin was not significant (P>0.05). However, the bonding strength of total etch groups was reduced significantly (P<0.05) after aPDT using both photosensitizers. Phycocyanin self-etch group showed the greatest bonding strength (19.48 ± 3.89 MPa) in comparison to other aPDT groups. No significant reduction after thermocycling in dentin bond strength was observed in all groups (P>0.05), with exception of control total etch groups, which showed significant difference (P<0.05). Conclusion: aPDT using toluidine blue O and phycocyanin has no adverse effect on bonding to sound dentin when using universal adhesive in self-etch protocol. Clinical relevance: Phycocyanin is more recommended to be used in aPDT in comparison to toluidine blue as a natural photosensitizer. [ABSTRACT FROM AUTHOR]

Periodontitis is an infectious disease characterized by the destruction of supporting tissues. Antimicrobial photodynamic therapy (aPDT) has been proposed as an improved method for eliminating microorganisms. Its efficiency depends on the correct use of physical and chemical parameters. Thus, these parameters and their relations were evaluated in this study with the purpose of establishing lethal conditions for combating bacterial agents. Diode lasers and light-emitting diodes (LEDs) were characterized to evaluate the absorption profile and resonance of methylene blue (MB) and toluidine blue O (TBO). The relations between light energy density and photosensitizer absorption were determined. Two methodologies were used to evaluate the effects of aPDT against Aggregatibacter actinomycetemcomitans. LED light exhibited a broad emission spectrum with a peak light wavelength of 637 nm and 99% purity. The resonance intensity of MB was higher with diode laser irradiation, and TBO showed higher resonance intensity with LED irradiation. There was no difference in the absorption profile of photosensitizers using diode lasers or LEDs, and variations in power density did not result in an increasing or decrease in light absorption. A. actinomycetemcomitans was susceptible to photodynamic processes. Emission spectra and peak light wavelengths of light sources combined with the absorption profiles of photosensitizers were the main parameters involved in determining the efficiency of photodynamic effects. Power density did not alter the light absorption of photosensitizers. The association between adequate irradiation characteristics and photosensitizer absorption results in complete inactivation of A. actinomycetemcomitans. In addition, the bactericidal effect was not altered by an increase in energy densities. [ABSTRACT FROM AUTHOR]

The study evaluates the dependence of p53 protein expression on the appearance of dark neurons (DNs) in the hippocampus in rats during experimental modeling of septoplasty. Septoplasty simulation was carried out on 15 sexually mature male Wistar rats. We studied histological sections of the hippocampus stained with Nissl toluidine blue and antibodies to the p53 protein. In the CA1 subfield, the number of p53-positive neurons significantly increased on the 2nd, 4th (p < 0.001) and 6th days (p < 0.05). In the dynamics, the peak of the growth of p53 protein expression in the cytoplasm of CA1 and CA2 neurons fell on the 2–4th day after the operation, and on the 6th day the number of these neurons decreased (p < 0.001). In the cytoplasm of CA3 neurons in all periods after surgery, an increase in the expression of the p53 protein as compared to the control group was noted. In the CA1 pyramidal layer, the number of DNs decreased on the 6th day (p < 0.001). In CA2, after 2 days, a minimum of DNs as compared with the 4th day (p < 0.001) was noted. In CA3, on the 4th day, there was a peak in DNs as compared with the rest of the days (p < 0.001). A positive strong association was found in all periods of assessment and in all subfields of the hippocampus between an increase in the number of dark and p53-positive neurons. The appearance of dark and p53-positive neurons in the hippocampal formation in rats after simulating septoplasty are typical responses of nervous tissue to stress. It is obvious that the expression of the p53 protein is associated with the basophilia of the cytoplasm of neurons, their morpho-functional state. Presumably, the p53 protein can trigger not only the activation of damaged neurons in the hippocampus but also play a neuroprotective role. Upcoming studies should determine the role of the p53 protein in the further fate of damaged neurons in the pyramidal layer and differentiate the mechanisms of its expression. [ABSTRACT FROM AUTHOR]

Spartium junceum L. is a typical species of Mediterranean shrubland areas, also grown in gardens and parks as an ornamental. In recent years in Europe, S. junceum has been recurrently found to be infected by different subspecies and genotypes of the quarantine regulated bacterium Xylella fastidiosa (Xf). This work presents for the first time the anatomy of S. junceum plants that we found, by means of genetic and immunochemistry analysis, to be naturally infected by Xf subsp. multiplex ST87 (XfmST87) in Monte Argentario (Grosseto, Tuscany, Italy), a new outbreak area within the EU. Our anatomical observations showed that bacteria colonized exclusively the xylem conductive elements and moved horizontally to adjacent vessels through pits. Interestingly, a pink/violet matrix was observed with Toluidine blue staining in infected conduits indicating a high content of acidic polysaccharides. In particular, when this pink-staining matrix was observed, bacterial cells were either absent or degenerated, suggesting that the matrix was produced by the host plant as a defense response against bacterial spread. In addition, a blue-staining phenolic material was found in the vessels and, at high concentration, in the pits and inter-vessels. SEM micrographs confirmed that polysaccharide and phenolic components showed different structures, which appear to be related to two different morphologies: fibrillary and granular, respectively. Moreover, our LM observations revealed bacterial infection in xylem conductive elements of green shoots and leaves only, and not in those of other plant organs such as roots and flowers. [ABSTRACT FROM AUTHOR]

Diabetic foot infection caused by multidrug-resistant bacteria, is becoming serious problem. Moreover, polymicrobial biofilms contribute significantly to the persistent infections. In the present study, we investigated the effectiveness of novel toluidine blue conjugated chitosan coated gold–silver core–shell nanoparticles (TBO–chit–Au–AgNPs) mediated photodynamic therapy and demonstrate their use as a nontoxic antibacterial therapy to combat diabetic foot ulcer (DFU) caused by multi-drug resistant strains both in monomicrobial and polymicrobial state of infection. In vitro efficacy of TBO–chit–Au–AgNPs mediated photodynamic therapy (PDT) against polymicrobial biofilms was determined using standard plate count method and compared with that of monomicrobial biofilms of each species. Different anti-biofilm assays and microscopic studies were performed to check the efficacy of TBO–chit–Au–AgNPs mediated PDT, displayed significant decrease in the formation of biofilm. Finally, its therapeutic potential was validated in vivo type-2DFU. Cytokines level was found reduced, using nano-phototheranostic approach, indicating infection control. Expression profile of growth factors confirmed both the pathogenesis and healing of DFU. Hence, we conclude that TBO–chit–Au–AgNPs mediated PDT is a promising anti-bacterial therapeutic approach which leads to a synergistic healing of DFU caused by MDR bacterial strains. [ABSTRACT FROM AUTHOR]

This study aimed to assess the validity and efficacy of blue dye in colposcopic assessment of genital injury in pre- and postmenopausal women with and without history of consensual sexual intercourse. Two hundred women were prospectively enrolled and examined colposcopically with and without toluidine blue dye in order to detect and categorize genital lesions (laceration, bruise and abrasion). Examination of genital trauma was accomplished in a standardized way and findings were photo documented. A wide range of influencing factors with a potential impact on prevalence and nature of genital injury was recorded beforehand using a questionnaire. The frequency of diagnostic injury differed substantially depending on the examination technique, ranging from 9% using colposcopic magnification only to 28% with the additional use of toluidine blue dye. A vertical laceration affecting the posterior fourchette was the most frequent lesion detected (17%, n = 32). Menopausal status seems to have significant impact on genital injury prevalence (p = 0.0165), as 42% (16/ 38) of postmenopausal compared to 24% (36/ 151) of premenopausal women had at least one genital lesion. Furthermore, vaginal medication (p = 0.0369), vaginal dryness (p = 0.0228), dyspareunia (p = 0.0234) and low frequency of sexual intercourse (p = 0.0022) were found to significantly correlate with the presence of genital lesions. According to our findings, standardized colposcopy in combination with toluidine blue dye facilitates accurate assessment of genital lesions. Genital trauma situated at another site than the posterior part of the vaginal introitus seems to be uncommon after consensual intercourse. [ABSTRACT FROM AUTHOR]

A label-free electrochemical aptasensor is reported for sensitive detection of the 6-kDa early secreted antigenic target (ESAT-6). For the first time, the bimetallic organic framework (b-MOF) of Zr-MOF-on-Ce-MOF was decorated with nitrogen-doped graphene (NG) and applied as the matrix for electroactive toluidine blue (Tb) to form the NG@Zr-MOF-on-Ce-MOF@Tb nanohybrid. The prepared nanohybrid with excellent hydrophilicity, dispersibility, and large specific surface exhibited significant electrochemical response. This nanohybrid could be directly used for anchoring ESAT-6 binding aptamers (EBA) through the interaction between the 5′-phosphate group (PO43−) of EBA and Zr4+ of Zr-MOF. The signal response before and after incubating the ESAT-6 antigen has been evaluated by cyclic voltammetry at a scan rate of 100 mV s−1 from − 0.7 to 0.3 V (vs. SCE). Under optimal conditions, the proposed aptasensor displayed a wide linear range from 100 fg mL−1 to 10 ng mL−1 with a limit of detection (LOD) of 12 fg mL−1. The developed method showed good reproducibility with a relative standard deviation (RSD) of 2.27%. The aptasensor showed favorable results in the analysis of the real samples. With these merits, the aptasensor has exceptional potential as a diagnostic tool for tuberculosis in clinical practice. [ABSTRACT FROM AUTHOR]

The electrochemical behavior of a phenothiazine dye, toluidine blue, in a gelatin hydrogel in the presence of trypsin, peroxidase, and their combination was studied by cyclic voltammetry. It has been shown that peroxidase contained in the hydrogel has a significant effect on the electrochemical transformations of toluidine blue during the first 30 s of measurements. The addition of trypsin enhances the effect of peroxidase catalysis. The possibility of simultaneous assessment of proteolysis and redox equilibrium in a model hydrogel system when analyzing current–voltage curves of cyclic voltammetry was experimentally justified. The results we obtained substantiate the scheme and principles of bioelectrochemical modeling of the diffusion of electroactive molecules in the extracellular matrix. [ABSTRACT FROM AUTHOR]

The surface carboxyl and sulfate groups on cellulose nanowhiskers were quantified via the adsorption of toluidine blue O (TBO), a cationic dye. Here, simple and rapid protocols, such as mixing the nanowhisker suspensions with a dye solution, separating the supernatants via centrifugation, and determining the excess dye concentration via visible light absorbance techniques, were used to obtain reproducible results comparable with those obtained via titration. In addition to facilitating the discrete quantification of the sulfate and carboxyl groups, the TBO adsorption method enabled carboxyl quantification in the presence of mercapto groups, which was difficult to achieve via titration. The adsorption of TBO onto the carboxyl groups was completed within 30 min, enabling the rapid treatment of many samples within a short period. [ABSTRACT FROM AUTHOR]

Development of a new pH sensor system, which is simple to prepare, sensitive, selective and workable with low volume, is demanding research in biomedical and environmental studies. In the literature, organic molecules like methylene blue, toluidine blue, hydroquinone, catechol, anthraquinone and polyaniline-based redox probes have been widely used for this purpose. In general, these redox probes have easily interfered with common biochemicals such as dopamine, ascorbic acid, NADH, H2O2, cysteine, hydrazine, and some transition metal ions, etc., and in turn to marked potential and current drifts (pH-false positive response). In this work, a highly redox-active, stable and interference-free redox polymer based on poly(4-chloroaniline) (PANI(4-Cl)) modified graphitized mesoporous carbon (GMC), designated as GMC@PANI(4-Cl), has been prepared using 4-chloroaniline as a monomer in pH 7 phosphate buffer solution. The new redox polymer system showed a distinct redox peak at Eo'= 0.15 V vs Ag/AgCl with a stable voltammetric response. Transmission electron microscope analysis of the redox polymer composite had shown adhesion of black polymeric solid due to polyaniline like the molecular system as a surface layer on the GMC material. The constructed calibration plot was linear in the pH window 2-11 with a slope and regression values − 58 mV pH−1 and 0.9997, respectively. The GMC@PANI(4-Cl) modified electrode showed a sensitive and selective pH monitoring without any interference from the common biochemicals as listed above. As a practical application, pH sensing of commercial pH solutions, undiluted urine and saliva samples were demonstrated. Also, a three-in-one screen-printed carbon modified GMC@PANI(4-Cl) was explored for pH monitoring of a bacterial (E.coli) growth, which showed a comparable response with the conventional pH electrode. [ABSTRACT FROM AUTHOR]

Introduction and hypothesis: As a notable chronic disorder, the incidence of interstitial cystitis (IC) has been documented to have increased among the female population with activity in microRNA-495 (miR-495) implicated in this disease. The current study was aimed at elucidating the effects associated with miR-495 on the inflammatory response and bladder fibrosis in rats with ulcerative IC via the JAK–STAT pathway by targeting JAK3. Methods: Ulcerative IC rat models were established. The targeting relationship between JAK3 and miR-495 was evaluated using luciferase reporter assay. After gain- or loss-of-function assays, mast-cell infiltration was assessed using toluidine blue staining, bladder fibrosis using Masson staining, and NO content using nitrate reductase method. JAK3 protein expression was detected by immunohistochemistry, JAK3, STAT1, STAT3, TGFβ-1, Col-I, Col-III, JAK1, JAK2, p-STAT1, and p-STAT3 expression by RT-qPCR and Western blot analysis, and serum IL-6, IL-8, IL-10, IL-17, and TNF-α levels in rats by ELISA. Results: Following transfection of overexpressed miR-495 or siRNA-JAK3, a diminished degree of mast-cell infiltration, number of mast cells, bladder fibrosis, NO content, JAK3-positive expression, mRNA expression of JAK3, STAT1, STAT3, TGFβ-1, Col-I, Col-III, protein expression of JAK1, JAK2, JAK3, p-STAT1, p-STAT3, and expression of IL-6, IL-8, IL-10, IL-17, and TNF-α were identified. Conclusions: Collectively, our key findings provide evidence supporting the notion that the overexpression of miR-495 ameliorates inflammatory response and bladder fibrosis in ulcerative IC rat models via inactivation of the JAK–STAT signaling pathway by inhibiting JAK3. [ABSTRACT FROM AUTHOR]

Objectives: The aim of this study was to investigate if bone regeneration can be promoted by homologous transplantation of STRO-1 sorted (STRO-1+) porcine tooth germ mesenchymal stem cells (TGSCs) with the combination of polyethylenglycol (PEG)-based hydrogel and biphasic calcium phosphate (BCP) scaffolds. Material and methods: TGSCs were isolated from impacted third molars of domestic pigs. Nine critical-sized defects were created as (1) untreated defect; filled with (2) autogenous bone; (3) BCP + PEG; (4) BCP + PEG + unsorted TGSCs; (5) BCP + unsorted TGSCs; (6) BCP + PEG + STRO-1-sorted TGSCs; (7) BCP + STRO-1-sorted TGSCs; (8) BCP + PEG + osteogenic induced unsorted TGSCs; and (9) BCP + PEG + osteogenic induced STRO-1-sorted TGSCs in 20 domestic pigs. CM-DiI labelling was used to track cells in vivo. Histomorphometric assessment of new bone formation was achieved by toluidine blue O staining and microradiography after 1, 2, 4 and 12 weeks posttransplantation. Results: Complete healing was achieved in all defects although defects with PEG hydrogel presented better bone formation while STRO-1+ and unsorted TGSCs showed similar ability to form new bone after 12 weeks. Transplanted cells were seen in defects where PEG hydrogel was used as carriers in contrast to defects treated with cells and only bone grafts. Conclusions: PEG hydrogel is an efficient carrier for homologous stem cell transplantation. TGSCs are capable of promoting bone healing in critical-sized defects in combination with bone graft and PEG hydrogel. Clinical relevance: This study provides information about the importance of the delivery vehicle for future translational stem cell delivery approaches. [ABSTRACT FROM AUTHOR]

Background: Vanilla planifolia is an important tropical orchid for production of natural vanilla flavor. Traditionally, V. planifolia is propagated by stem cuttings, which produces identical genotype that are sensitive to virulent pathogens. However, propagation with seed germination of V. planifolia is intricate and unstable because the seed coat is extremely hard with strong hydrophobic nature. A better understanding of seed development, especially the formation of impermeable seed coat would provide insights into seed propagation and conservation of genetic resources of Vanilla. Results: We found that soaking mature seeds in 4% sodium hypochlorite solution from 75 to 90 min significantly increased germination. For the culture of immature seeds, the seed collection at 45 days after pollination (DAP) had the highest germination percentage. We then investigated the anatomical features during seed development that associated with the effect of seed pretreatment on raising seed germination percentage. The 45-DAP immature seeds have developed globular embryos and the thickened non-lignified cell wall at the outermost layer of the outer seed coat. Seeds at 60 DAP and subsequent stages germinated poorly. As the seed approached maturity, the cell wall of the outermost layer of the outer seed coat became lignified and finally compressed into a thick envelope at maturity. On toluidine blue O staining, the wall of outer seed coat stained greenish blue, indicating the presence of phenolic compounds. As well, on Nile red staining, a cuticular substance was detected in the surface wall of the embryo proper and the innermost wall of the inner seed coat. Conclusion: We report a reliable protocol for seed pretreatment of mature seeds and for immature seeds culture based on a defined time schedule of V. plantifolia seed development. The window for successful germination of culturing immature seed was short. The quick accumulation of lignin, phenolics and/or phytomelanins in the seed coat may seriously inhibit seed germination after 45 DAP. As seeds matured, the thickened and lignified seed coat formed an impermeable envelope surrounding the embryo, which may play an important role in inducing dormancy. Further studies covering different maturity of green capsules are required to understand the optimal seed maturity and germination of seeds. [ABSTRACT FROM AUTHOR]

The present work deals with the synthesis of a novel quaternary photocatalyst ZrCdPbO4, prepared by co-precipitation method in controlled conditions. The prepared material was characterized using XRD, IR, FESEM, EDX, UV–VIS and XPS analysis to verify the crystalline nature, functional, morphological, chemical composition, band gap energy and oxidation states of elements. Generation of hydroxyl radicals was confirmed by the quenching experiment. These radicals were further used for degradation of some organic pollutants [Toluidine Blue (TB), Brilliant Green (BG) and Crystal Violet (CV)] and maximum degradation conditions like pH, concentration of dye, amount of photocatalyst and intensity of light were extracted by kinetic study. The maximum degradation rate was observed for CV = 10.8 × 10−4 (s−1), for TB = 8.62 × 10−4 (s−1) and for BG = 5.37 × 10−4 (s−1). [ABSTRACT FROM AUTHOR]

The concentration level of cytokeratin fragment antigen 21-1 (CYFRA21-1) can be used as an important indicator for predicting non–small cell lung cancer (NSCLC). Here, a sandwich-type electrochemical immunosensor for ultrasensitive detection of CYFRA21-1 is developed. The sensor based on a combination of gold nanoparticle (AuNPs) decorated Ti3C2Tx-MXene (Au-Ti3C2Tx) as the substrate enhancer, and toluidine blue (TB) modified AuNPs doped covalent organic framework (COF) polymer as the signal tag (TB-Au-COF). The Au-Ti3C2Tx is used to capture numerous primary antibodies and accelerate the electron transfer rate of the substrate, while the TB-Au-COF can be applied to provide a large number of signal units TB and secondary antibodies. These features of composites endow the proposed immunosensor with high sensitivity and current response to CYFRA21-1. Under optimum conditions, the immunosensor offers a wide current response for CYFRA21-1 from 0.5–1.0 × 104 pg·mL−1 with a detection limit of 0.1 pg·mL−1. Furthermore, the biosensing platform can be applied for CYFRA21-1 detection to analyze real serum samples, providing an effective and useful avenue for the applicability of Au-Ti3C2Tx and TB-Au-COF composite materials in biosensing field. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the effects of toluidine blue-mediated photodynamic therapy (TB-PDT) on the periodontitis-induced bone resorption in periodontitis in rats. Periodontal disease was induced by cotton ligature around the right second maxillary molar in 64 rats. After 4 weeks, the rats were randomly divided into four groups: sterile saline solution (control group); laser therapy (laser group); TB (100 μg/mL); TB plus laser (0.15 W/cm2) irradiation every other day for 240 s (PDT group). All rats were euthanized at 15 days postoperatively. Eight gingival tissue samples were collected from each group. The expressions of receptor activator of nuclear factor kappa-Β ligand (RANKL) and osteoprotegerin (OPG) in gingival tissue samples were detected by real-time quantitative polymerase chain reaction (RT-qPCR). The maxillae from the rest of the rats were taken for histological examination. In the PDT group, the analysis revealed less bone loss than in the control treatment (P < 0.05). No significant difference was found among the control group, TB group, and laser group (P > 0.05). Significantly higher and lower expressions of RANKL and OPG were revealed in the PDT group than that in control group, respectively (P < 0.01). When compared with the control group, the expression of RANKL was significantly reduced by 40.0% in periodontitis in rats treated with TB-PDT for 15 days (P < 0.01). The expression of OPG was increased in the PDT group with TB-PDT for 15 days, when compared with the control group (P < 0.05). TB-PDT treatment significantly reverses the abnormal expression of RANKL and OPG in periodontitis in rats. [ABSTRACT FROM AUTHOR]

In this work, iron terephthalate metal–organic framework (MOF-235) as an efficient adsorbent has been synthesized and employed for toluidine blue (TB) removal in a batch system. The adsorbent was characterized by Fourier-transform infrared, scanning electron microscopy, and powder X-ray diffraction. The effects of operational parameters including pH, initial dye concentration (Cd), and the adsorbent dosage (m) on the TB adsorption were studied. A Box–Behnken design (BBD) was applied to identify the effective factors on the removal efficiency (R%) of toluidine blue. The optimal conditions were as follows: m = 0.02 g, Cd = 100 mg L−1, and pH = 4.5. The equilibrium adsorption isotherms and kinetic models were also studied. Since the results of the Langmuir model fitted well to all the experimental data (R2 = 0.9972), the maximum monolayer adsorption of TB was 180.4 mg g−1. Furthermore, the pseudo-second-order model describes well the process. Iron terephthalate metal–organic framework (MOF-235) shows a great potential to remove TB from aqueous solution. [ABSTRACT FROM AUTHOR]

Objective: The in vivo mechanoresponsive and lubricating changes of the mandibular condylar cartilage (MCC) associated with mandibular lateral shift (MLS) and recovery are poorly understood. Using growing rats, we investigated whether the expression of mechanoresponsive factors, including proteoglycan-4 (PRG4), Indian hedgehog (Ihh) and transforming growth factor-β1 (TGF-β1), would be affected by MLS. We also investigated whether these changes could recover to the control level after a 2-week treatment reversal (TR). Materials and methods: The MLS appliances were placed for 2 or 4 weeks in 5-week-old rats and removed from 7-week-old rats in the TR group. The MCC was analysed histomorphometrically by toluidine blue staining. Reverse transcription-polymerase chain reaction and immunohistochemistry were performed to evaluate the expression of PRG4, Ihh, PTHrP (parathyroid hormone-related protein), TGF-β1, Matrix metallopeptidase 13 (MMP-13) and a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS-5). Results: A thickened superficial layer and an enhanced expression of PRG4 were detected in MLS groups. PTHrP-Ihh expression correlated positively with the up-regulation of PRG4. TGF-β1 expression decreased in the early stage of MLS but recovered to the control level in the TR group. A significantly enhanced expression of MMP-13 in MLS groups was detected. Conclusion: MLS treatment, which acted on the growth stage of rats, affected the morphology and expression of lubrication factor in the MCC. Elimination of this mechanical stimulus may help MCC recover to normal conditions. Clinical relevance: Our study supports that the adaptive changes of MCC, which are caused by mandibular functional deviation, could be largely recovered by early treatment. [ABSTRACT FROM AUTHOR]